Antibacterial and Anti-anthracnose Activities of Endophytic Fungi Colonized in Blumea balsamifera (L.) DC.

doi:10.11924/j.issn.1000-6850.casb2020-0501

Abstract

Abstract:

To obtain abundant endophytic fungi from Blumea balsamifera (L.) DC., and select bio-control strains for discovery of bioactive metabolites, water-agar method was used to isolate endophytic fungi from Blumea balsamifera (L.) DC distributed in Hainan Island. ITS analysis was adopted for identification and analysis of endophytic fungi. Furthermore, the activities of fermented endophytes culture extract against four strains of bacteria were tested by filtering paper method. Plate confrontation was carried out to evaluate the inhibitory effects of endophytes on four strains of anthracnose. As a result, 191 strains were isolated and identified, which belonged to 27 genuses, 45 species, the dominant genuses included Diaporthe, Paraphoma, Ectophoma, Penicillium, Talaromyces, Hypoxylon, Phomopsis, Colletotrichum and Fusarium. Clonostachys, Fusarium and Diapothe were more active in antibacterial assay. Anti-anthracnose results showed that the endophyts belonged to Talaromyces, Ectophoma and Penicillium toughly inhibited four strains of anthracnose. In conclusion, our research preliminary discussed the category variety of endophytic fungi derived from Blumea balsamifera collected in Hainan Island, bioactivity-guided screening was carried out firstly, and the selected active endophytic fungi may serve as candidates for founding new agronomic agents.

Key words: Blumea balsamifera (L.) DC., endophytic fungi, antimicrobial activity, anti-anthracnose activity, microbial agent

CLC Number:

S482.2+91

Yuan Chao, Shu Xuechun, Zhang Yingbo, Wang Kai, Xie Xiaoli, Xu Ziqi, Yuan Yuan. Antibacterial and Anti-anthracnose Activities of Endophytic Fungi Colonized in Blumea balsamifera (L.) DC.[J]. Chinese Agricultural Science Bulletin, 2021, 37(23): 38-44.

Figures/Tables 5

References 25

[1]	韦睿斌, 庞玉新, 杨全, 等. 艾纳香黄酮类化学成分研究进展[J]. 广东药学院学报, 2014, 30(1):123-127.
[2]	De B H J, Hugo J, Cotingting C, et al. Medicinal plants for women’s healthcare in southeast Asia: A meta-analysis of their traditional use, chemical constituents, and pharmacology[J]. Journal of Ethnopharmacology, 2014, 151(2):747-767. doi: 10.1016/j.jep.2013.11.030 pmid: 24269772
[3]	邹婧, 杨全, 庞玉新, 等. 艾纳香口腔护理液的处方筛选及其抑菌效果研究[J]. 广东药学院学报, 2015, 31(5):571-575.
[4]	Wang Y H, Yu X Y. Biological Activities and Chemical Compositions of Volatile Oil and Essential Oil from the Leaves of Blumea balsamifera[J]. Journal of essential oil-bearing plants JEOP, 2018, 21(6):1511-1531.
[5]	Aly A H, Debbab A, Prokech P. Fungal endophytes-secret producers of bioactive plant metabolites[J]. Die Pharmazie-An International Journal of Pharmaceutical Sciences, 2013, 68(7):499-505.
[6]	Zhang H W, Song Y C, Tan R X. Biology and chemistry of endophytes[J]. Natural Product Reports, 2006, 23(5):753-771. pmid: 17003908
[7]	Gao H, Li G, Lou H X. Structural diversity and biological activities of novel secondary metabolites from endophytes[J]. Molecules, 2018, 23(3):646. doi: 10.3390/molecules23030646 URL
[8]	Li G, Lou H X. Strategies to diversify natural products for drug discovery[J]. Medicinal Research Reviews, 2018, 38(4):1255-1294. doi: 10.1002/med.2018.38.issue-4 URL
[9]	唐青, 周思旋, 雷帮星, 等. 贵州西南部艾纳香内生真菌多样性研究[J]. 菌物学报, 2017, 36(11):1498-1503.
[10]	元超, 郭玉华, 张影波, 等. 艾纳香内生真菌Corynespora cassiicola J9次生代谢产物研究[J]. 药学学报, 2019, 54(5):892-896.
[11]	元超, 李刚, 张影波, 等. 艾纳香内生真菌Diaporthe sp.次生代谢产物[J]. 微生物学报, 2018, 59(4):753-761.
[12]	Yuan C, Yang H X, Guo Y H, et al. New α-pyrones from an endophytic fungus, Hypoxylon investiens J2[J]. RSC Adevances, 2019, 9:27419-27423.
[13]	Kusari S, Lamshoft M, Zuklke S, et al. An endophytic fungus from Hypericum perforatum that produces hypericin[J]. Journal of Natural Products, 2008, 71(2):159-162. doi: 10.1021/np070669k URL
[14]	Guo Y H, ShuX C, Zhang Y B, et al. Screening of bioactive secondary metabolites of endophytes associated with Blumea balsamifera (L.) DC. by UPLC-QTof-MS, DNP and bio-assay [J/OL]. Microbiol China, 2019:1-14.
[15]	Kumar S, Stecher G., Li M, et al. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms[J]. Molecular Biology and Evolution, 2018, 35(6):1547-1549. doi: 10.1093/molbev/msy096 URL
[16]	范翠, 卢永仲, 康冀川, 等. 毛筒壳科真菌次级代谢产物生物活性的评价[J]. 菌物学报, 2019, 38(4):560-574.
[17]	LI G, Kusari S, Lamshoft M, et al. Antibacterial secondary metabolites from an endophytic fungus, Eupenicillium sp. LG41[J]. Journal of Natural Products, 2014, 77(11):2335-2341. doi: 10.1021/np500111w URL
[18]	周而勋, 杨媚. 从植物病组织中分离立枯丝核菌的快速、简便技术[J]. 华南农业大学学报, 1998, 19(1):125-126.
[19]	何美莹, 徐荣. 橡胶种子、橡胶种子油的营养及食用安全性研究[J]. 热带农业科技, 2013, 36(3):1-6,10.
[20]	李君, 兰国玉, 李玉武. 海南儋州橡胶林与热带次生林土壤微生物区系研究[J]. 热带作物学报, 2016, 37(3):433-438.
[21]	张逸飞, 刘娟娟, 孟磊, 等. 农业利用对海南省天然次生林土壤微生物的影响[J]. 生态学报, 2015, 35(21):6983-6992.
[22]	杨娟, 董醇波, 陈万浩. 等不同地区杜仲树皮内生真菌群落组成及生态功能结构的差异分析[J]. 中国中药杂志, 2019, 44(6):1126-1134.
[23]	Gamboa M A, Laureano S, Bayamn P. Measuring diversity of endophytic fungi in leaf fragments: Does size matter?[J]. Mycopathologia, 2003, 156(1):41-45. doi: 10.1023/A:1021362217723 URL
[24]	Konstantinos M, Land M L, Brettin T S. The fast changing landscape of sequencing technologies and their impact on microbial genome assemblies and annotation[J]. Plos One, 2012, 7(12):48837. doi: 10.1371/journal.pone.0048837 pmid: 23251337
[25]	张爱梅, 韩雪英, 王嘉等. 马衔山中国沙棘根瘤内共生细菌多样性研究[J]. 生态学报, 2019, 39(1):294-301.

菌株目/属	组织部位	菌株数	相对多度	备注	菌株目/属	组织部位	菌株数	相对多度	备注
Diaporthe	根茎叶	18	9.42	优势菌群	Cladosporium	茎叶	5	2.62
Acrocalymma	叶	2	1.05		Daldinia	茎叶	3	1.57
Paraphoma	茎叶	8	4.19	优势菌群	Hypoxylon	茎叶	9	4.71	优势菌群
Alternaria	叶	2	1.05		Xylariaceae	茎叶	2	1.05
Phoma	茎叶	2	1.05		Nigrospora	茎叶	6	3.14
Boeremia	叶	1	0.52		Pestalotiopsis	叶	3	1.57
Ectophoma	茎叶	11	5.76	优势菌群	Muscodor	叶	2	1.05
Pleosporaceae	根茎叶	1	0.52		Phomopsis	茎叶	16	8.38	优势菌群
Corynespora	叶	2	1.05		Colletotrichum	根茎叶	22	11.52	优势菌群
Periconia	叶	2	1.05		Clonostachys	根	4	2.09
Pleosporales	叶	3	1.57		Fusarium	根茎叶	33	17.28	优势菌群
Penicillium	根	11	5.76	优势菌群	Inaequalispora	根	4	2.09
Talaromyces	根茎叶	15	7.85	优势菌群	Trichoderma	根	1	0.52
Diutina	叶	3	1.57

菌株目/属	组织部位	菌株数	相对多度	备注	菌株目/属	组织部位	菌株数	相对多度	备注
Diaporthe	根茎叶	18	9.42	优势菌群	Cladosporium	茎叶	5	2.62
Acrocalymma	叶	2	1.05		Daldinia	茎叶	3	1.57
Paraphoma	茎叶	8	4.19	优势菌群	Hypoxylon	茎叶	9	4.71	优势菌群
Alternaria	叶	2	1.05		Xylariaceae	茎叶	2	1.05
Phoma	茎叶	2	1.05		Nigrospora	茎叶	6	3.14
Boeremia	叶	1	0.52		Pestalotiopsis	叶	3	1.57
Ectophoma	茎叶	11	5.76	优势菌群	Muscodor	叶	2	1.05
Pleosporaceae	根茎叶	1	0.52		Phomopsis	茎叶	16	8.38	优势菌群
Corynespora	叶	2	1.05		Colletotrichum	根茎叶	22	11.52	优势菌群
Periconia	叶	2	1.05		Clonostachys	根	4	2.09
Pleosporales	叶	3	1.57		Fusarium	根茎叶	33	17.28	优势菌群
Penicillium	根	11	5.76	优势菌群	Inaequalispora	根	4	2.09
Talaromyces	根茎叶	15	7.85	优势菌群	Trichoderma	根	1	0.52
Diutina	叶	3	1.57

内生真菌	马铃薯炭疽菌	西瓜炭疽菌	艾纳香炭疽菌	香蕉炭疽菌
Clonostachys rosea	++	++	++	++
Corynespora cassiicola	+	++	++	++
Diutina catenulata	-	+	+	+
Ectophoma pomi	+++	+++	++	++
Paraphoma chrysanthemicola	++	+	+	+
Penicillium sp.	+++	++	++	++
Phomopsis sp.	+	+	+	+++
Diaporthe phaseolorum	+	++	++	++
Fusarium equiseti	++	++	++	+
Muscodor sp.	+	+	+	++
Periconia byssoides	+	+	+	++
Pestalotiopsis microspora	+	+	++	++
Pleosporaceae sp.	+	++	+	++
Talaromyces pinophilus	++	+++	+++	++

内生真菌	马铃薯炭疽菌	西瓜炭疽菌	艾纳香炭疽菌	香蕉炭疽菌
Clonostachys rosea	++	++	++	++
Corynespora cassiicola	+	++	++	++
Diutina catenulata	-	+	+	+
Ectophoma pomi	+++	+++	++	++
Paraphoma chrysanthemicola	++	+	+	+
Penicillium sp.	+++	++	++	++
Phomopsis sp.	+	+	+	+++
Diaporthe phaseolorum	+	++	++	++
Fusarium equiseti	++	++	++	+
Muscodor sp.	+	+	+	++
Periconia byssoides	+	+	+	++
Pestalotiopsis microspora	+	+	++	++
Pleosporaceae sp.	+	++	+	++
Talaromyces pinophilus	++	+++	+++	++

内生真菌	铜绿假单胞菌	大肠埃希菌	金黄色葡萄球菌	枯草芽孢杆菌
Clonostachys rosea	+++	+	+++	+++
Corynespora cassiicola	-	-	++	-
Cladosporium herbarum	-	-	+	-
Diutina catenulata	+	-	++	+
Daldinia sp.	-	-	+	-
Diaporthe phaseolorum	+++	+	+++	+++
Fusarium equiseti	+++	-	+++	+++
Nigrospora hainanensis	++	+	+	++
Penicillium sp.	++	-	++	++
Phomopsis sp.	++	+	++	++
Pestalotiopsis microspora	++	+	+	++
Pleosporaceae sp.	+	-	+	-
Talaromyces pinophilus	+	+	-	+
Trichoderma sp.	++	+	++	++
Inaequalispora prestonii	++	+	+++	+
CK	+++	+++	++	++